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Richard Stelluto

Introducing a New Source for Endotoxin-free Water

Introducing a New Source for Endotoxin-free Water

Prepared Biologicals offers a wide range of packaging sizes, from 100 ml for a pilot applications to 200 L for biomanufacturing applications

Contamination of cell culture has long been a major problem both for biological  research and manufacturing applications and can arise from multiple sources, such as bacteria, viruses, and fungi. Among them, endotoxin is by far the most difficult to control. Endotoxin is a complex lipopolysaccharide (LPS) which is present on an outer membrane of most gram-negative bacteria, with as many as 2 million copies per single cell, and is shed during both the growth and especially the cell death. The effects of endotoxin on the cell culture varies considerably, depending on the cell type, mechanism of action, and applications. In established cell lines, endotoxin sensitivity is highly variable which may affect downstream applications (1-3). In contrast, other cell lines (4) stem cells (5), primary cells (6), and especially those involved in the immune response such as dendritic cells (7), may be sensitive to low levels of endotoxin. Although not fully known, the mechanisms by which endotoxin affects the cells includes the interactions with the cell surface receptors such as CD14, endocytosis, and binding to the cytoskeleton (reviewed in 1). The consequences of endotoxin contamination could include: lower recombinant protein yield, reduced transfection efficiency, lower cell yield, and presence of undesirable cell population during the cell derivation from the hematopoetic stem cell progenitors. Furthermore, low levels of endotoxins can cause the systemic shock and death in the patients.  As the therapeutic use of cell culture-derived biologics, such as recombinant proteins, antibodies, and cells rapidly expands, endotoxin-free cell culture is becoming critical to success.

Despite the challenges posed by the ubiquitousness of endotoxin in the environment, there are several ways to control endotoxin levels in cell culture. These include: working under aseptic conditions, using low-endotoxin sera and other media additives (8), baking glassware at 180-250 0C in addition to autoclaving, and/or ensuring the plasticware is certified to be endotoxin-free (1). Most importantly, water used for media preparation and equipment washing has to be endotoxin- free. It is critical to prepare the water using well-maintained equipment, and keep it endotoxin-free by storing it in well-irradiated and properly sealed containers. In many cases, producing endotoxin-free water in-house on a consistent basis is impractical, cost-wise.  In those cases, endotoxin-free water can be purchased from commercial suppliers, which could also be an expensive alternative.

To meet the market needs for affordable, cell culture certified endotoxin-free water, Prepared Biologicals offers a wide range of packaging sizes, from 100 ml needed for a pilot experiment to as much as 200 L for biomanufacturing applications.  A secured lot option is available.

To place an order or for more information, visit http://preparedbiologicals.com/reagents/, call 800-767-0766, or fax 617-997-4525. 

About Prepared Biologicals

Prepared Biologicals, LLC is a manufacturer of premium biological products including a complete line of Endotoxin-Free biological buffers. We uniquely specialize in custom formulations, OEM manufacturing and offer Secured Lot Reagents for those experiments that demand a Single Reagent Lot for the entire course of an experiment. Our special packaging sizes include 55 gallon drums (200L) which are available for commercial and Bioprocess applications. The goal of Prepared Biologicals is to meet the changing needs of the scientific community with innovative products that ensure the best possible experimental outcome.

Company contact:

 

Prepared Biologicals, LLC

Tel:

1-800-767-0766

Email:

customercare@preparedbiologicals.com

 

References:

  1. Ryan, J. A. 2004. Endotoxins and cell culture, p. 1-8. In Corning Life Sciences Technical Bulletin. http://www.corning.com/lifesciences/technical_information/techdocs/endotoxincc.asp.
  2. Weber, M., Möller, K., Welzcek, M., Schorr, J. 1995. Effects of lipopolysaccharide on transfection efficiency in eukaryotic cells. BioTechniques 19: 931-939.
  3. Butash, K.A., Natarajan, P., Young, A., Fox, D.K. 2000. Reexamination of the Effect of Endotoxin on Cell Proliferation and Transfection Efficiency. Biotechniques 29: 610-619.
  4. Rieder, A., Grimmer, S., Aachmann F.L., Westereng B., Kolset, S.O., Knutsen, S.H. 2013. Generic Tools to Assess Genuine Carbohydrate Specific Effects on In Vitro Immune Modulation Exemplified by -glucans. Carbohydrate Polymers 92: 2075– 2083.
  5. Nolte J. http://www.orfgenetics.com/media/PDF/University_of_Gottingen_flyer.pdf
  6. Unger, R.E., Peters, K., Sartoris, A., Freese C., Kirkpatrick, C. J. 2014. Human Endothelial Cell-Based Assay for Endotoxin as Sensitive as the Conventional Limulus Amebocyte Lysate Assay. Biomaterials 35 (10): 3180-3187
  7. Schwarz, H., Schmittner, M., Duschl, A., Horejs-Hoeck, J. 2014. Residual Endotoxin Contaminations in Recombinant Proteins Are Sufficient to Activate Human CD1c+ Dendritic Cells. PLoS ONE 9(12): e113840. doi:10.1371/journal.pone.0113840
  8. Impact of Media Additives on Stem Cell Behavior in Culture. https://www.mti-globalstem.com/image/catalog/Application/Impact%20of%20Media%20Additives%20on%20Stem%20Cell%20Behavior%20in%20Culture.pdf